A method for shortening agarose gel electrophoresis staining
An agarose gel and agarose technology, which is applied in the field of biochemical experiments, can solve the problems of difficult degradation, transformation and purification, cumbersome, harmful, toxic and other problems, and achieves an effect that is beneficial to observation.
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Embodiment 1
[0032] Such as figure 1 Shown is the gel electrophoresis image extracted by the method of the present invention.
[0033] A method for shortening the staining of agarose gel electrophoresis, which comprises: cleaning and drying the graduated cylinder, the triangular flask, the gel plate, and the tooth sample lattice, sealing the both sides of the opening of the gel plate with adhesive tape, and Place the gel tray on a horizontal table, insert the tooth sample grid, and use the graduated cylinder to measure the TBE electrophoresis buffer.
[0034] Since the tooth sample grid has a flat opening on one side and a notch on the other side, it is a complementary design to the gel tray. When inserting, you need to pay attention to the left and right directions.
[0035] Preparation of agarose gel
[0036]
[0037] Use a measuring cylinder to measure 40mL of 5×TBE electrophoresis buffer, add water to 200mL, configure 1×TBE to dilute the electrophoresis buffer and pour it into a c...
Embodiment 2
[0047]Example 2. Prior art traditional EB added directly to the gel
[0048] EB-stained gel electrophoresis was figure 2 .
[0049] Preparation of DNA Molecular Weight Ladders
[0050] The lambda DNA fragment obtained by EcoR I or Hind III enzyme was used as the molecular weight standard during electrophoresis. λDNA is a double-stranded DNA molecule with a length of about 50kb. The concentration of its commercial solution is 0.5mg / mL. The enzymatic hydrolysis operation is as follows:
[0051] HindⅢ cut DNA to obtain 8 fragments, the lengths were 23.1, 9.4, 6.6, 4.4, 2.3, 2.0, 0.56 and 0.12kb.
[0052] After cutting IDNA with EcoRI, six fragments were obtained, the lengths of which were 21.2, 7.4, 5.8, 5.6, 4.9 and 2.5kb.
[0053] Preparation of agarose gel
[0054] Take 20mL of 5×TBE buffer and add water to 200mL to prepare 0.5×TBE dilution buffer for use.
[0055] Preparation of glue solution: Weigh 0.4g agarose, put it in a 200mL conical flask, add 0.5×TBE dilution bu...
Embodiment 3
[0065] Example 3. Prior art improved nucleic acid agarose gel electrophoresis EB staining method
[0066] Improved nucleic acid agarose gel electrophoresis such as image 3 shown.
[0067] Dissolve 1g of agarose in 100mL of TAE buffer, heat it in a microwave oven, and when it cools down to about 50°C, add 5uLEB stock solution to the agarose gel to prepare a piece of 1% agarose gel without EB. 5uL EB stock solution was recorded as EBO in 95uL distilled water, and then 1uL was taken out of it and added to 1mL of bromophenol blue to mix, which was recorded as EBO1. When loading the sample, the former directly takes 5uL of the PCR product and 3uL of bromophenol blue without EB and mixes the sample, while the latter fully mixes 5uL of the PCR product and 3uL of bromophenol blue (EBO1) containing EB and loads the sample, both under the same conditions Perform electrophoresis, electrophoresis at 100V for 15min, and detect the brightness of the bands under ultraviolet light.
[006...
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